1. Field of the Invention
The present invention relates to a cantilever for measuring intra-cellular and inter-cellular microspaces suitable for measurement or operation in intra-cellular and inter-cellular microspaces.
2. Related Art Statement
In recent years of bio-field, there has been focused on elucidating or controlling cellular vital functions.
Such elucidation or control of cellular vital functions involves essential operations such as measuring electric potential difference in intra-cellular and inter-cellular microspaces and introducing and taking a gene or the like into and out of a cell.
A number of suggestions have been made on method or device for introducing into a cell a substance to be introduced such as a gene. Known examples of such suggestions include disclosures in Japanese Unexamined Patent Publication Nos. 2003-325161 and 2006-166884.
The Japanese Unexamined Patent Publication No. 2003-325161 describes a suggestion on a cell operating device and a cell operating method using the cell operating device. This publication discloses a technique on a configuration which is capable of detailed real-time observation of serial dynamics changes shown on the cell during an interval from introduction to manifestation of a gene, which is applicable even for elucidating or controlling cell differentiation, and which further includes a probe of an Atomic Force Microscope (AFM) which is provided, as cell operative means capable of minimizing cellular damage during introduction of a gene or the like, with a needle-shaped matter for inserting into a cell a substance relating to gene or gene manifestation which is fixed thereto.
However, the technique described in the Japanese Unexamined Patent Publication No. 2003-325161 has a problem in that the substance relating to gene or gene manifestation remains fixed to the needle-shaped matter, so that the gene or the like is not constantly held in the cell when the needle is pulled out, resulting in incapability of producing a gene recombinant.
A proposal in view of such problem is made in Japanese Unexamined Patent Publication No. 2006-166884, which describes a proposal on a method to introduce a substance into a cell.
The Japanese Unexamined Patent Publication No. 2006-166884 discloses a technique regarding a method for introducing into a cell a substance to be introduced, by binding the substance to be introduced to a needle-shaped material by means of an intermediate binding force such that the substance to be introduced is caused to remain in the cell when the needle-shaped material is punctured thereinto. This publication also discloses a technique relating to a configuration in which an atomic force microscope (AFM) is used for a method of detecting puncture of the needle-shaped material into the cell and as an introducing device for introducing the substance to be introduced into the cell.
Specifically, in the method described in the Japanese Unexamined Patent Publication No. 2006-166884, as shown in FIG. 21, a probe 101 provided to an AFM cantilever 100 is etched by a focused ion beam so as to be configured as a needle-shaped material, and further, the probe 101 as the needle-shaped material is used to perform a puncturing operation into a cell 103 and a nucleus 104 in the cell 103.
The Japanese Unexamined Patent Publication No. 2006-166884 also discloses a technique related to electrostatic coupling as a method to couple the substance to be introduced to the needle-shaped material by means of an intermediate binding force. Because such electrostatic coupling is a weak coupling, the substance to be introduced can be easily separated in the cell. In electrostatic coupling, cationic polymer is generally used.
Specifically, in the method described in the Japanese Unexamined Patent Publication No. 2006-166884, as shown in a schematic block diagram of FIG. 22, a thiol group is introduced into a needle-shaped material 105, which is a silicon-made AFM probe etched to have a sharpened edge, to modify succinimide active ester on a surface of needle-shaped material to immobilize polylysine followed by electrostatic coupling of plasmid phrGFP as the substance to be introduced, before introducing the needle-shaped material 105 into the cell.
However, the technique described in the Japanese Unexamined Patent Publication No. 2006-166884 has a problem in that the electrostatic coupling, which is weak, can be canceled, resulting in the substance to be introduced to be detached before introduction into the cell, thus making it impossible to introduce a desired material into the cell.
Further, the techniques described in the Japanese Unexamined Patent Publication No. 2006-166884 suggests or discloses nothing in terms of taking a substance out of a cell and measuring intra-cellular electric potential distribution. Accordingly, there is desired a specific device that allows stable intra-cellular measurement or operation to be easily performed.
The present invention has been made in view of the above-described problems, and an object of the present invention is to provide a cantilever for measuring intra-cellular and inter-cellular microspaces that allows stable intra-cellular measurement or operation to be easily performed, which is durable, reliable, and low cell-invasive.
Another object of the present invention is to provide a cantilever for measuring intra-cellular and inter-cellular microspaces that allows performing, easily and stably with good reproducibility and minimum variation, nano-cell mapping, i.e., measurement in intra-cellular and inter-cellular microspaces and introducing and taking a substance to and out of the microspaces.